Understanding human pancreas development for diabetes cell-replacement therapy

阐明调控早期胰腺发育的关键因素对于我们长期追求理解疾病机制和开发基于人多能干细胞（hPSC）的糖尿病β细胞替代疗法至关重要。然而，胰腺特化所需的信号线索和转录因子的完整景观仍不清楚。利用hPSC定向分化和CRISPR/Cas介导的基因组编辑，Huangfu实验室最近发表的工作确定了人类胰腺分化过程中RFX 6，GATA 6和GATA 4（已知与新生儿和成人发病糖尿病相关的基因）的关键新要求。在未发表的工作中，我们进一步发现了胰腺分化中对FOXA 2的一种新的剂量依赖性需求，FOXA 2是一种与糖尿病和高胰岛素血症相关的基因。在Huangfu，Leslie和Pe'er实验室之间的合作中，我们将采用互补的遗传学，基因组学和计算方法，并利用hPSC分化来解剖人类胰腺发育。我们将利用遗传学方法建立精确的hPSC疾病模型，并研究疾病表型背后复杂的遗传相互作用，并采用基因组学方法，包括ChIP-seq和ATAC-seq分析以及尖端的单细胞转录组学，以了解人类胰腺发育和β细胞功能的调节因子。我们的发现将增强对人类胰腺发育和疾病的理解，并促进改进的hPSC定向分化方案的开发，用于产生用于疾病研究和治疗的功能性β细胞。

Delineating key factors that regulate early pancreas development is crucial to our long-term pursuit of understanding disease mechanisms and developing human pluripotent stem cell (hPSC) based β-cell replacement therapies for diabetes. However, a complete landscape of signaling cues and transcription factors required for pancreas specification remains unclear. Using hPSC directed differentiation and CRISPR/Cas-mediated genome editing, recently published work from the Huangfu lab has identified critical new requirements for RFX6, GATA6 and GATA4 (genes known to be associated with neonatal and adult-onset diabetes) during human pancreatic differentiation. In unpublished work, we have further uncovered a novel, dose-dependent requirement for FOXA2, a gene associated with diabetes and hyperinsulinism, in pancreatic differentiation. Here in this collaboration between the Huangfu, Leslie and Pe’er labs, we will undertake complementary genetic, genomic and computational approaches and utilize hPSC differentiation to dissect human pancreatic development. We will utilize genetic approach to create precise hPSC disease models and interrogate complex genetic interactions underlying disease phenotypes, and employ genomic approaches including ChIP-seq and ATAC-seq analyses and cutting-edge single-cell transcriptomics to understand regulators of human pancreatic development and β cell function. Our findings will enhance the understanding of human pancreatic development and disease, and facilitate the development of improved hPSC directed differentiation protocols for the generation of functional β cells for disease study and treatment.